The invention belongs to the field of biotechnology and particularly relates to the screening and application of bacillus licheniformis MEL09 with a high 3-hydroxy butanone yield. A bacillus licheniformis strain is preserved in China Center for Type Culture Collection (CCTCC) with the collection number of CCTCC NO:M209173. The biological characteristics of the strain are that: the strain is rod-shaped; a bacterial colony on an LB solid culture medium is round, has irregular edges and is capillary; and a lawn is dry, non-transparent, pervasive and in grey white color, produces spores and is positive in a V.P. reaction. Glucose of the strain is taken as a carbon source, and 40.66g/L 3-hydroxy butanone can be produced by performing culturing at the temperature of 37 DEG C for 36 hours. The 3-hydroxy butanone produced by the strain is an important chemical raw material and can be widely applied to fields such as food, chemical industry, medicament and the like.